ISOLATION, IDENTIFICATION, AND QUANTIFICATION OF LECTIN PROTEIN CONTENTS IN CHAMERION ANGUSTIFOLIUM L. DRIED RAW MATERIAL AND THE STUDY OF ITS ACTIVITY USING RATUSERYTROAGGLUTINATION
Keywords:
Lectins, Chamerion angustifolium L, leaves, Buds, Flowers, RatuserytroagglutinationAbstract
Objective: To isolate, identify and quantify lectin protein contents in Chamerion angustifolium L. dried raw material species namely; leaves in the flowering stage, buds, and flowers and the study of its activity.
Methods: Lectin activity has been determined using the biological method called ratuserytroagglutination. This method is based on the formation of aggregates of lectins and rat erythrocytes using the floor amount of lectins that agglutinate erythrocytes as a unit of measurement. Additionally, the protein contents of the extracts have been determined using the Bradford assay method.
Results: lectins activity from Chamerion angustifolium L. in leaves in the flowering stage, buds, and flowers were 2.72±0.06, 0.24±0.008, and 0.56±0.014 units/mg protein, respectively. The greatest lectins activity was in the leaves in the flowering stage followed by flowers and then in buds. Protein contents in leaves in the flowering stage, buds, and flowers were 4.71±0.03, 6.77±0.02, and 5.76±0.14 mg/mL, respectively.
Conclusion: All proteins obtained from the Chamerion angustifolium L. plant raw material were shown to possess rat erythrocytes agglutinating activity. The crude extract of leaves in the flowering stage exhibited the strongest hemagglutinating activity of about 2.72 units/mg proteins, whereas the buds showed the lowest activity of about 0.24 units/mg proteins. It should be highlighted herein, although many plant lectins mimic the behavior of plant storage proteins, these lectins should not be classified as storage proteins.
Â
Downloads
References
Zündorf I. Teedrogen und phytopharmaka von max wichtl (Hrsg.). Pharm Unserer Zeit 2009;38:192–203.
Maruška A, Ragažinskiene O, Vyšniauskas O, Kaškoniene V, Bartkuviene V, Kornyšova O, et al. Flavonoids of willow herb (Chamerion angustifolium (L.) Holub) and their radical scavenging activity during vegetation. Adv Med Sci 2014;59:136–41.
Kiss A, Kowalski J, Melzig MF. Compounds from Epilobium angustofolium inhibit the specific metallopeptidases ACE, NEP and APN. Planta Med 2004;70:919–23.
Vitalone A, McColl J, Thome D, Costa LG, Tita B. Characterization of the effect of Epilobium extracts on human cell proliferation. Pharmacology 2003;69:79–87.
Lebeda AF, Jurenko NI, Isaikina AP, Soko VG. Medicinal plants: the fullest encyclopedia. Moscow: AST-Press Book; 2004.
Vitalone A, Bordi F, Baldazzi C, Mazzanti G, Saso L, Tita B. Antiproliferative effect on a prostatic epithelial cell line (PZ-HPV-7) by Epilobium angustifolium L.. Il Farmaco 2001;56:483–9.
Schepetkin IA, Kirpotina LN, Jakiw L, Khlebnikov AI, Blaskovich CL, Jutila MA, et al. Immunomodulatory activity of oenothein B isolated from Epilobium angustifolium. J Immunol 2009;183:6754–66.
KaÅ¡koniene V, StankeviÄius M, Drevinskas T, Akuneca I, KaÅ¡konas P, Bimbiraite-Surviliene K, et al. Evaluation of phytochemical composition of fresh and dried raw material of introduced Chamerion angustifolium L. using chromatographic, spectrophotometric and chemometric techniques. Phytochemistry 2015;115:184–93.
Moilanen J, Sinkkonen J, Salminen JP. Characterization of bioactive plant ellagitannins by chromatographic, spectroscopic and mass spectrometric methods. Chemoecology 2013;23:165–79.
Battinelli L, Tita B, Evandri MG, Mazzanti G. Antimicrobial activity of Epilobium spp. Extracts. Il Farmaco 2001;56:345–8.
Kiss AK, Bazylko A, Filipek A, Granica S, Jaszewska E, Kiarszys U, et al. Evaluation of phytochemical composition of fresh and dried raw material of introduced Chamerion angustifolium L. using chromatographic, spectrophotometric and chemometric techniques. Phytochemistry 2015;115:184–93.
Ruszova E, Cheel J, Pávek S, Moravcova M, Hermannová M, Matějková I, et al. Epilobium angustifolium extract demonstrates multiple effects on dermal fibroblasts in vitro and skin photoprotection in vivo. Gen Physiol Biophys 2013;32:347–59.
Rubinstein N, Ilarregui JM, Toscano MA, Rabinovich GA. The role of galectins in the initiation, amplification and resolution of the inflammatory response. Tissue Antigens 2004;64:1–12.
Ye XY, Ng TB, Tsang PWL, Wang J. Isolation of a homodimeric lectin with antifungal and antiviral activities from red kidney bean (Phaseolus vulgaris) seeds. J Protein Chem 2001;20:367–75.
Abdullaev FI, de MejÃa EG. Antitumor effect of plant lectins. Natural Toxins 1997;5:157–63.
Pusztai A, Grant G, Spencer RJ. Kidney bean lectin-induced Escherichia coli overgrowth in the small intestine is blocked by GNA, a mannose-specific lectin. J Appl Bacteriol 1993;75:360–8.
Barrientos LG, Gronenborn AM. The highly specific carbohydrate-binding protein cyanovirin-N: structure, anti-
HIV/Ebola activity and possibilities for therapy. Mini Rev Med Chem 2005;5:21–31.
Pollicita M, Schols D, Aquaro S, Peumans WJ, Van Damme EJM, Perno CF, et al. Carbohydrate-binding agents (CBAs) inhibit HIV-1 infection in human primary monocyte-derived macrophages (MDMs) and efficiently prevent MDM-directed viral capture and subsequent transmission to CD4+T lymphocytes. Virology 2008;370:382–91.
Wimer BM. Characteristics of PHA-L4, the mitogenic isolectin of phytohemagglutinin, as an ideal biologic response modifier. Molecular Biotherapy J 1990;2:4–17.
Kiran KK, Lalith PCK, Sumanthi J, Sridhar RG, Chandra SP, Reddy BVR. The biological role of lectins. J Orofacial Sci 2012;4:20–5.
Ann, MH. Role of lectins (and rhizobial exopolysaccharides) in legume nodulation. Curr Opin Plant Biol 1999;2:320–6.
Balaji P, Arunk P, Senthil KR, Meenakshi SM, Brindha P. Isolation and identification of an adaptogenic protein from Cicer arietinum Linn. Int J Pharm Pharm Sci 2012;4:79–82.
Yufang H, Yubao H, Liu Y, Guang Q, Jichang L. Extraction and purification of a lectin from red kidney bean and preliminary immune function studies of the lectin and four chinese herbal polysaccharides. J Biomed Biotechnol 2010:1–9. doi:10.1155/ 2010/217342. [Epub 2010 Oct 03]
Abhinav KV, Sharma A, Vijayan M. Identification of mycobacterial lectins from genomic data. Proteins 2013;81:644–57.
Athamna A, Cohen D, Athamna M, Ofek I, Stavri H. Rapid identification of Mycobacterium species by lectin agglutination. J Microbiol Methods 2006;65:209–15.
Arishya S, Tzi BN, Jack HW, Peng L. Purification and characterization of a lectin from Phaseolus vulgaris cv. (Anasazi Beans). J Biomed Biotechnol 2009:1–9. doi:10.1155/ 2009/929568. [Epub 2009 Mar 25]
Rybak L, Rudik G. Research on the quantitative content of lectins in plants of the Geranium L. Genus. Pharma Innovation 2013;2:38–41.
Smillie TJ, Khan IA. A comprehensive approach to identifying and authenticating botanical products. Clin Pharmacol Ther 2010;87:175–86.
Abudeiyh ZH, Sereda PI, Karpiuk UV, Lutenko IA. The study of anatomy and numeric indices of the aerial parts of Chamerion angustifolium (L.) Holub. Farmacom 2011;3:39–43.
Pogorila NF, Surzhyk LM, Pogorila ZO. A new method of plant lectin is testing. Ukr Biokhim Zh 2002;59:217–20.
Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248–54.
State Pharmacopoeia of the USSR; 1987. p. 336S.
Rozhnova NA, Gerashchenkov GA, Babosha AV. The effect of arachidonic acid and viral infection on the phytohemagglutinin activity during the development of tobacco acquired resistance. Russ J Plant Physiol 2003;50:661–5.